Cell manufacturing on an autonomous platform

Abstract

A method for cell culture expansion and maintenance, comprising seeding, in a first cell culture cassette, a plurality of cell colonies adhered to a first surface of the first cell culture cassette; capturing, by an optical engine, time-series images of the plurality of cell colonies as the plurality of cell colonies expand; determining, by a platform manager, a subset of cell colonies to remove from the first surface based on the time-series images; removing, by the optical engine, the subset of cell colonies from the first surface; washing, by a fluid management system, the removed subset of cell colonies from the first cell culture cassette; repeating steps for one or more iterations; harvesting a set of remaining cell colonies from the first cell culture cassette; seeding the set of remaining cell colonies into a second cell culture cassette; repeating steps for additional iterations in the second cell culture cassette.

1 . A method of cell culture management, comprising:
 (a) seeding, in a first cell culture cassette, a plurality of cell colonies adhered to a first surface of the first cell culture cassette;   (b) capturing, by an optical engine, time-series images of the plurality of cell colonies as the plurality of cell colonies expand;   (c) determining, by a platform manager, a subset of cell colonies to remove from the first surface based on the time-series images;   (d) removing, by the optical engine, the subset of cell colonies from the first surface;   (e) washing, by a fluid management system, the removed subset of cell colonies from the first cell culture cassette;   (f) repeating steps (b)-(e) for one or more iterations;   (g) harvesting, by the optical engine, a set of remaining cell colonies from the first cell culture cassette;   (h) seeding the set of remaining cell colonies into a second cell culture cassette; and   (i) repeating steps (b)-(g) for one or more iterations for the second cell culture cassette.   
     
     
         2 . The method of  claim 1 , further comprising:
 (j) repeating steps (h)-(i) for one or more additional cell culture cassettes.   
     
     
         3 . The method of  claim 2 , further comprising harvesting cells for use in a cell therapy after the completion of step (j). 
     
     
         4 . The method of  claim 1 , wherein determining the subset of cell colonies comprises applying a machine learning model to the time-series images. 
     
     
         5 . The method of  claim 1 , wherein determining the subset of cell colonies comprises ranking the plurality of cell colonies according to a set of criteria and removing one or more lowest ranked cell colonies. 
     
     
         6 . The method of  claim 5 , wherein the set of criteria comprises parameters corresponding to clonal colony quality. 
     
     
         7 . The method of  claim 5 , wherein determining the subset of cell colonies comprises removing a portion of a highest ranked cell colony for analysis. 
     
     
         8 . The method of  claim 1 , wherein determining the subset of cell colonies comprises removing cell colonies or portions of cell colonies to reduce cell confluence on the first surface. 
     
     
         9 . The method of  claim 1 , wherein determining the subset of cell colonies comprises removing cell colonies or portions of cell colonies to shape remaining cell colonies in a pattern. 
     
     
         10 . The method of  claim 1 , wherein harvesting the first set of remaining cell colonies comprises harvesting a single, clonal cell colony. 
     
     
         11 . The method of  claim 1 , wherein harvesting the set of remaining cell colonies comprises harvesting all cells therein when cell confluence reaches a predetermined threshold. 
     
     
         12 . The method of  claim 1 , wherein the set of remaining cell colonies is transferred into the second cell culture cassette as single cells, cell colonies, or cell sheets. 
     
     
         13 . The method of  claim 1 , wherein the first surface comprises an optical film configured for imaging and optical removal of cells by the optical engine. 
     
     
         14 . The method of  claim 1 , wherein steps (b)-(f) are performed over a period of at least 10 days. 
     
     
         15 . The method of  claim 1 , wherein determining the subset of cell colonies comprises analyzing morphological characteristics of the plurality of cell colonies in the time-series images. 
     
     
         16 . The method of  claim 15 , wherein the morphological characteristics include at least one of colony size, colony shape, cell density within the colony, or cell arrangement within the colony. 
     
     
         17 . The method of  claim 1 , wherein the optical engine comprises a laser-based cell removal tool configured to selectively remove cells from the first surface. 
     
     
         18 . The method of  claim 1 , wherein seeding the set of remaining cell colonies into the second cell culture cassette comprises dissociating the remaining cell colonies into single cells prior to seeding. 
     
     
         19 . The method of  claim 1 , further comprising applying a rejuvenation protocol to the cells in the second cell culture cassette. 
     
     
         20 . The method of  claim 19 , wherein the rejuvenation protocol comprises introducing factors for partial cellular reprogramming. 
     
     
         21 . A system for cell culture management, comprising:
 a first cell culture cassette, configured for a plurality of cell colonies to be seeded to and adhere to a first surface thereof;   an optical engine configured to capture time-series images of the plurality of cell colonies as the plurality of cell colonies expand;   a platform manager configured to determine a subset of cell colonies to remove from the first surface based on the time-series images, the optical engine being further configured to remove the subset of cell colonies from the first surface;   a fluid management system configured to wash the removed subset of cell colonies from the first cell culture cassette; and   a second cell culture cassette configured for a set of remaining cell colonies harvested from the first cell culture cassette to be seeded therein.   
     
     
         22 . The system of  claim 21 , configure to perform a method according to any one of  claims 1-20 . 
     
     
         23 . A computer-implemented method of cell culture management, comprising:
 receiving time-series images of a plurality of cell colonies adhered to a first surface of a first cell culture cassette as the plurality of cell colonies expand;   determining a subset of cell colonies to remove from the first surface based on the time-series images;   sending instructions to an optical engine to remove the subset of cell colonies from the first surface;   sending instructions to a fluid management system to wash the removed subset of cell colonies from the first cell culture cassette;   sending instructions to the optical engine to harvest a set of remaining cell colonies from the first cell culture cassette; and   sending instructions to the fluid management system to seed the set of remaining cell colonies into a second cell culture cassette.   
     
     
         24 . A computer program product for cell culture management, the computer program product comprising a computer readable storage medium having program instructions embodied therewith, the program instructions executable by a processor to cause the processor to perform a method according to  claim 23 .